Significant improvements in the treatment of cystic fibrosis (CF) have occurred over the past 30 years due to drug delivery via inhalation aerosols. However, the efficacy inhaled therapies is dramatically reduced in CF patients because of the presence of a viscous mucus transport barrier within the airways, extensive degradation and metabolism of inhaled drug prior to exerting its pharmacological action, and the development mucoid Pseudomonas aeruginosa colonies. Often drugs or gene vectors cannot reach the intended target before their activity has been reduced or eliminated. Poor transport efficiencies in drug delivery have lead to the failure of therapies including the inability to attain the relatively low efficiencies required for gene therapy success. Applying the unique transport and active properties of nanoscale systems in drug delivery is a promising strategy for overcoming the biological barriers in CF lung disease. In particular, the ability of nanoparticles systems to exert strong influences on their environment using heat (a nanoknife) and magnetic fields (nano-pullies) are attractive functional attributes for increasing transport and drug distribution in CF lung disease. The long term objectives are to overcome these barriers and achieve critical improvements in CF therapy. The CENTRAL HYPOTHESIS of the proposed research is that novel multifunctional nanoparticles will facilitate significant enhancement of the efficacy of model therapeutic agents due to increased diffusion and penetration through mucus and biofilm barriers in cystic fibrosis when administered as an aerosol. Our preliminary data have demonstrated that (1) marked increases in particle and bulk transport of nanoparticles can be attained during nano-pulley mode, (2) we prove that our nanoparticles can be heated and they act like nanoknives cutting through biopolymers such as DNA that causes CF mucus to be diffusion limiting, (3) we demonstrate also our ability to synthesize a number of different types of magnetic nanoparticles for use in these studies: core-shell composites for use in image studies and surface functionlized particles for drug conjugation (4) we then functionalized these particles, attaching a model drug to the surface using a bio- cleavable conjugation scheme, (5) Drug release could be triggered using magnetic fields, (6) and finally, we then loaded the nanoparticles into inhalable microparticles suitable for aerodynamic lung targeting. These exciting preliminary data strongly support the rationale and feasibility of the proposed approach. Moreover, the interdisciplinary team has a very strong record in each of the critical areas of the project: pulmonary drug delivery, nanoparticles design and engineering, and the molecular genetics and microbiology of CF. The main objective of the proposed research is to develop, synthesize, characterize, and evaluate novel particle systems that simultaneously allow controlled lung deposition and enhanced transport in CF disease. These systems will provide high drug concentrations delivered directly to the site of action and will therefore facilitate significant improvements in drug and gene therapies in CF, prolonging survival and enhancing quality of life. Therefore, the SPECIFIC AIMS of the exploratory R21 phase of the project are (i) Prepare and characterize multifunctional nanoparticles and incorporate them into micro-systems suitable for inhalation via a dry powder aerosolization, (ii) Characterize drug transport and delivery performance of particles for CF therapy in vitro, and (iii) Evaluate in vivo efficacy and safety of the multifunctional nanoparticle pulmonary delivery system versus aerosolized controls. The results of the R21 phase will demonstrate the initial feasibility of this approach. The R33 phase of the proposed work will be focused on optimization of the nanosystems and biocompatibility evaluation. This innovative application of nanotechnologies to CF lung disease will also have potential in other lung diseases where transport barriers to drug delivery exist: tuberculosis, lung cancer, COPD, etc. The proposed studies will fill important gaps in our understanding of the systems of pulmonary drug delivery of nanosystems and subsequently how controlled administration of drug and gene therapies may impact CF treatment strategies.